ChIP-Atlas: SRX9702043

Antigen

Cell type Class: Blood
Cell type: Macrophages
MeSH Description: The relatively long-lived phagocytic cell of mammalian tissues that are derived from blood MONOCYTES. Main types are PERITONEAL MACROPHAGES; ALVEOLAR MACROPHAGES; HISTIOCYTES; KUPFFER CELLS of the liver; and OSTEOCLASTS. They may further differentiate within chronic inflammatory lesions to EPITHELIOID CELLS or may fuse to form FOREIGN BODY GIANT CELLS or LANGHANS GIANT CELLS. (from The Dictionary of Cell Biology, Lackie and Dow, 3rd ed.)

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: For histone ChIP-seq, lysates were clarified from MNase digested nuclei and histone-DNA complexes were isolated with H3K4me1,H3K4me3 or H3K27Ac antibodies. For p300 and Hdac3 ChIP-seq, lysates were clarified from sonicated nuclei and p300-DNA or Hdac-DNA complexes were isolated with p300 or Hdac3 antibodies. Illumina libraries were constructed using a modified custom paired-end protocol. In brief, samples were end-repaired (1*T4 DNA ligase buffer, 0.4 mM dNTP mix, 2.25 U T4 DNA polymerase, 0.75 U Klenow DNA polymerase and 7.5 U T4 polynucleotide kinase; 30 min at 21–25 C), A-tailed (1*NEB buffer 2, 0.4 mM dNTPs and 3.75 U of Klenow (exo-); 30 min at 37 C) and ligated (1*rapid DNA ligation buffer, 1 mM Illumina PE adapters and 1,600 U DNA ligase; 1–8 h at 21–25 C). Ligated fragments were amplified using indexed primers (Illumina) for 8–12 PCR cycles. DNA was purified with 1.8 volume KAPA DNA purification beads between each step.